Summary: The facultative methanol utilizer AM1 contains at least three genes ( and ) that encode functions involved in providing calcium to the holoenzyme of methanol dehydrogenase, the enzyme that oxidizes methanol to formaldehyde in this strain. Methane-utilizing bacteria (methanotrophs) also contain methanol dehydrogenase, and evidence suggests that similar methanol oxidation (Mox) functions may be present in some of these strains. DNA fragments from AM1 specific to and were isolated for use as hybridization probes against genomic digests of a variety of methanotrophic bacteria. Only the probe showed substantial hybridization, and it was used to identify and isolate an 8·5 kb dIII fragment from BG8 (a Type I methanotroph). Hybridization of restriction digests of this fragment to individual probes for AM1 and indicated that the relative gene order in BG8 is . A T7 dual promoter/polymerase protein expression system indicated that five polypeptides are expressed from a 4·5 kb region of BG8 DNA in , all transcribed in the same direction, and they apparently correspond to . The functions of and are currently not known, but the order of is reversed in BG8 compared to AM1. When subclones o the BG8 fragment containing these genes were used as hybridization probes to genomic digests of methanotrophic bacteria, specific bands were detected that suggested a similar gene order in most cases. These data indicate that the region is relatively highly conserved in methanotrophs, and that in most cases the genes are grouped together in the same order as in the facultative methanol utilizer .


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